Enzyme-linked immunosorbent assays for detection of equine antibodies specific to Sarcocystis neurona surface antigens.
Abstract: Sarcocystis neurona is the primary causative agent of equine protozoal myeloencephalitis (EPM), a common neurologic disease of horses in the Americas. We have developed a set of enzyme-linked immunosorbent assays (ELISAs) based on the four major surface antigens of S. neurona (SnSAGs) to analyze the equine antibody response to S. neurona. The SnSAG ELISAs were optimized and standardized with a sample set of 36 equine sera that had been characterized by Western blotting against total S. neurona parasite antigen, the current gold standard for S. neurona serology. The recombinant SnSAG2 (rSnSAG2) ELISA showed the highest sensitivity and specificity at 95.5% and 92.9%, respectively. In contrast, only 68.2% sensitivity and 71.4% specificity were achieved with the rSnSAG1 ELISA, indicating that this antigen may not be a reliable serological marker for analyzing antibodies against S. neurona in horses. Importantly, the ELISA antigens did not show cross-reactivity with antisera to Sarcocystis fayeri or Neospora hughesi, two other equine parasites. The accuracy and reliability exhibited by the SnSAG ELISAs suggest that these assays will be valuable tools for examining the equine immune response against S. neurona infection, which may help in understanding the pathobiology of this accidental parasite-host interaction. Moreover, with modification and further investigation, the SnSAG ELISAs have potential for use as immunodiagnostic tests to aid in the identification of horses affected by EPM.
Publication Date: 2005-09-09 PubMed ID: 16148170PubMed Central: PMC1235790DOI: 10.1128/CDLI.12.9.1050-1056.2005Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Antibodies
- Antigen
- Clinical Pathology
- Diagnosis
- Diagnostic Technique
- Disease Diagnosis
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Equine Diseases
- Equine Health
- Equine Protozoal Myeloencephalitis
- Horses
- Immune Response
- Immunology
- Infection
- Infectious Disease
- Parasites
- Pathogens
- Sarcocystis
- Serology
- Veterinary Medicine
- Veterinary Research
Summary
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This research study focuses on developing enhanced testing methods for detecting antibodies in horses related to Sarcocystis neurona, the main parasite causing equine protozoal myeloencephalitis. The study demonstrates the viability of enzyme-linked immunosorbent assays in accurately tracking equine immune response to the parasite which may be useful in diagnosing and understanding the disease.
Objective of the Research
- The main objective of this research was to develop an improved method of detecting equine antibodies specific to Sarcocystis neurona, the main cause of equine protozoal myeloencephalitis (EPM), a common neurological disease affecting horses in the Americas.
Proposed Method
- The researchers proposed the development of several enzyme-linked immunosorbent assays (ELISAs) based on the four major surface antigens of S. neurona (SnSAGs).
- The goal of these assays was to analyze the equine antibody response to S. neurona in a more standardized and reliable method than the current gold-standard of Western blotting against total S. neurona parasite antigen.
Results of the Study
- The study found the recombinant SnSAG2 (rSnSAG2) ELISA to have the highest sensitivity and specificity at 95.5% and 92.9%, respectively. This suggests that it could be an effective tool for diagnosing and studying S. neurona in horses.
- In contrast, the rSnSAG1 ELISA achieved only 68.2% sensitivity and 71.4% specificity, indicating this particular antigen may not be a reliable serological marker for analyzing antibodies against S. neurona in horses.
Significance of the Findings
- Importantly, the researchers found that the ELISA antigens did not cross-react with antisera to Sarcocystis fayeri or Neospora hughesi, two other equine parasites. This greatly improves the accuracy of the tests and their capacity to specifically detect S. neurona parasites.
- The improved accuracy and reliability of the SnSAG ELISAs mean they can potentially be valuable tools for examining the equine immune response against S. neurona infection. By doing so, these could help in understanding the biology of the accidental parasite-host interaction between S. neurona and horses.
- With some modification and further research, the SnSAG ELISAs could also be used as immunodiagnostic tests to aid in the identification of horses affected by EPM.
Cite This Article
APA
Hoane JS, Morrow JK, Saville WJ, Dubey JP, Granstrom DE, Howe DK.
(2005).
Enzyme-linked immunosorbent assays for detection of equine antibodies specific to Sarcocystis neurona surface antigens.
Clin Diagn Lab Immunol, 12(9), 1050-1056.
https://doi.org/10.1128/CDLI.12.9.1050-1056.2005 Publication
Researcher Affiliations
- Department of Veterinary Science, University of Kentucky, Lexington, KY 40546-0099, USA.
MeSH Terms
- Animals
- Antibodies, Protozoan / analysis
- Antibodies, Protozoan / blood
- Antibodies, Protozoan / cerebrospinal fluid
- Antibody Specificity
- Antigens, Surface / immunology
- Blotting, Western
- Cross Reactions
- Encephalomyelitis / diagnosis
- Encephalomyelitis / parasitology
- Encephalomyelitis / veterinary
- Enzyme-Linked Immunosorbent Assay / methods
- Horse Diseases / diagnosis
- Horse Diseases / parasitology
- Horses
- Recombinant Proteins / immunology
- Sarcocystis / immunology
- Sarcocystis / isolation & purification
- Sarcocystosis / diagnosis
- Sarcocystosis / immunology
- Sarcocystosis / veterinary
- Sensitivity and Specificity
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Citations
This article has been cited 4 times.- Alvarado-Esquivel C, Howe DK, Yeargan MR, Alvarado-Esquivel D, Alfredo Zamarripa-Barboza J, Dubey JP. Seroepidemiology of Sarcocystis neurona and Neospora hughesi infections in domestic donkeys (Equus asinus) in Durango, Mexico. Parasite 2017;24:27.
- Reed SM, Furr M, Howe DK, Johnson AL, MacKay RJ, Morrow JK, Pusterla N, Witonsky S. Equine Protozoal Myeloencephalitis: An Updated Consensus Statement with a Focus on Parasite Biology, Diagnosis, Treatment, and Prevention. J Vet Intern Med 2016 Mar-Apr;30(2):491-502.
- Dubey JP, Howe DK, Furr M, Saville WJ, Marsh AE, Reed SM, Grigg ME. An update on Sarcocystis neurona infections in animals and equine protozoal myeloencephalitis (EPM). Vet Parasitol 2015 Apr 15;209(1-2):1-42.
- Wendte JM, Miller MA, Nandra AK, Peat SM, Crosbie PR, Conrad PA, Grigg ME. Limited genetic diversity among Sarcocystis neurona strains infecting southern sea otters precludes distinction between marine and terrestrial isolates. Vet Parasitol 2010 Apr 19;169(1-2):37-44.
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